Project 1 - Chemistry & In Vitro Studies of Chinese Herbal Remedies

酗酒和吸毒在美国和世界各地造成了严重的医疗、社会和经济问题。为了开发有效的治疗方法，人们付出了大量的努力。不幸的是，在过去的40年里，只有三种疗效有限的药物获得了美国食品和药物管理局的批准。可卡因依赖的复杂性和缺乏有效的补救措施，特别是对复发的补救，这往往是 即使在长期禁欲之后，戒断和/或强烈的渴望也会引发这种症状，这对治疗构成了严峻的挑战。草药改变可卡因行为效应的一个可能的作用机制是通过kappa阿片受体。由于中国在18世纪和19世纪最初开发的许多草药疗法是针对鸦片成瘾的，因此将在项目1中筛选出化学成分 对于kappa受体活性，可以发现具有kappa激动剂或拮抗剂作用。已经证实，可卡因的一些滥用相关效应可以通过kappa阿片受体来改变，这些效应与kappa阿片类药物对伏隔核多巴胺水平的调节有关。例如，在大鼠中，kappa阿片激动剂已被证明可以减弱可卡因刺激的运动活动、可卡因辨别和可卡因自身给药。此外，kappa阿片激动剂也被证明可以减弱可卡因引起的伏隔核细胞外多巴胺水平的增加。此外，最近的研究还表明，kappa拮抗剂可能有助于防止阿片类药物和可卡因的复发。因此，财团致力于替代药物治疗将具有重要的临床意义，特别是在防止复发方面。 我们选择了两种中草药YGT(NPI-025)和XJL(NPI-028)用于这个项目，因为它们在治疗药物滥用方面的疗效在中国身上得到了证实，而且我们实验室近年来获得了令人鼓舞的科学数据。本研究项目一的具体目标是(1)采用高效液相指纹图谱技术对这两种中草药进行获取和标准化；(2)通过分级提供纯化的成分作为内标和分子 目的：(1)为阐明作用机制提供必要的工具；(3)将这些标准化的中草药及其部分分发到研究项目1、2和4，分别用于体外、体内和临床评估；以及(4)进行中草药及其部分的体外筛选，以确定其与u、Delta和kappa阿片受体、伤害素/孤儿FQ(N/OFQ)受体、D1和D2多巴胺受体的亲和力，以及通过[35S]GTPGammas结合和腺苷环化酶活性来研究它们的受体功能。在体外显示阳性结果的草药或部分将在项目2中使用各种体内模型进行检查。只有体内活性最好的才会接受项目4的临床评估。

Alcohol and drug abuse pose serious medical, social, and economic problems in the United States and around the world. A great deal of effort has been directed toward developing effective therapies. Unfortunately, in the last 40 years, only three medications with limited efficacy have been approved by the U.S. Food and Drug Administration. The complexity of cocaine dependence and the lack of effective remediation, especially for relapse, which is often precipitated by withdrawal and/or intense craving even after prolonged abstinence, poses a serious therapeutic challenge. One possible mechanism of action underlying the modification of cocaine's behavioral effects by herbal remedies is through kappa opioid receptors. Because many herbal remedies that were originally developed during the 18th and 19th centuries in China were targeted at opium addiction, chemical fractions that will be screened in Project 1 for kappa receptor activity may be found to have kappa agonist or antagonist actions. It is well established that some abuse related effects of cocaine can be modified through kappa opioid receptors, and these effects have been related to kappa opioid modulation of dopamine levels in the nucleus accumbens. For example, in rats, kappa opioid agonists have been shown to attenuate cocaine-stimulated locomotor activity, cocaine discrimination and cocaine self-administration. Moreover, kappa opioid agonists have also been shown to attenuate cocaine-induced increases in extracellular levels of dopamine in the nucleus accumbens. In addition, recent studies also indicate that kappa antagonists may be useful in preventing relapse to opioid and cocaine use. Therefore, a consortium effort focusing on alternative pharmacotherapies would have important clinical significance, especially in preventing relapse. We selected two Chinese herbal medicines, YGT (NPI-025) and XJL (NPI-028), for this project because of their proven efficacy in the treatment of substance abuse in China and encouraging scientific data obtained in recent years in our laboratories. The specific aims of this Research Project 1 are (1) to procure and standardize these two herbal medicines by HPLC fingerprinting; (2) to provide purified components by fractionation as internal standards and as molecular tools to elucidate the mechanism of action; (3) to distribute these standardized herbal medicines and fractions to Research Projects 1, 2, and 4 for in vitro, in vivo, and clinical evaluations respectively; and (4) to conduct in vitro screening of Chinese herbal remedies and fractions for their affinity at the mu, delta and kappa opioid receptors, the nociceptin / orphanin FQ (N/OFQ) receptor and D1 and D2 dopamine receptors and for their receptor functions by [35S]GTPgammaS binding and adenylyl cyclase activity. The herbal medicines or fractions that show positive results in vitro will be examined in Project 2 using various in vivo models. Only those with the best in vivo activity will be subiect to clinical evaluation in Project 4.